GUACAMOLE AND AVOCADO PRODUCTS

1. www.hiperbaric.comHiperbaric © 2013
GUACAMOLE AND AVOCADO
PRODUCTS
INTRODUCTION
High Pressure Processing (HPP) is a non-thermal food processing technology that allows
preserving nutrients, fresh taste and appearance of avocado and guacamole, with a longer
and safer shelf-life. On this sector of vegetable-derived products, the pressure used is
generally 600 MPa (6000 bar / 87000 psi) applied typically over 3 minutes at refrigerated
temperature.
Regarding to a physico-chemical effect on food, the HPP technology is softer than a thermal
treatment: it does not break or create covalent bonds, and does not create new compounds
by molecule degradation, such as in a conventional thermal process. However, HPP is able
to break, or create, weak bonds (hydrophobic and electrostatic interactions), only present
on macromolecules (Cheftel, 1992). It allows microorganism inactivation without modifying
the food nutritional quality and without significantly altering enzymatic activities. To
minimize the residual microorganism growth, the enzymatic reactions and changes in
sensory attributes, avocado products and guacamole must be stored at chilled temperature.
There are many reasons that make the HPP technology beneficial:
- Safer food products with a longer shelf-life are produced, thanks to the inactivation of
vegetative microorganisms (bacteria, yeasts and molds).
- Sensory food quality is maintained, keeping the fresh-like taste of home-made product.
- Nutritional quality is preserved.
- Suitable for “clean label” and organic food products.

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FOOD SAFETY AND LONGER SHELF-LIFE
Consumers have a keen interest in preservative-free food
products. HPP technology is an effective method to reduce
the microbiota without adversely affecting the sensory
attributes and decreases the dependence on additives
(Waite et al., 2009).
Guacamole is a minimally processed avocado product,
providing conditions that are ideal for the growth of
spoilage bacteria. Since these food products are ready to
eat, microbial growth must be strictly controlled.
Thermal treatments and chemical preservatives are the
traditional technologies used in preservation of these
products. These treatments reduce microbial load but lead
to an unacceptable sensory quality such as the generation
of bitter off-flavors (Jacobo-Velázquez and Hernández-
Brenes, 2010; López-Malo, et al., 1999).
Shelf- life increase
Guacamole spoils within the first 5 days, even when it is
stored at 5 ºC (41 ºF) (López-Malo et al., 1999). Lactic acid
bacteria, molds and yeasts are the main groups of
microorganisms responsible of the quick spoilage of these
product, thus their control by HPP should be a priority.
At 600 MPa (87,000 psi) held at for 3 min, significant
instantaneous reduction in microbial load of guacamole
(pH 6.35) was achieved without any previous acidification
(Jacobo-Velázquez and Hernández-Brenes, 2010). Levels of
mesophilic aerobic and lactic acid bacteria, two of the
spoilage indicators, remained constant during the first 40
days of storage at 4 ºC (39.2 ºF) in HPP guacamole, around
2 log cfu/g (Figure 1).
Molds and yeasts were well controlled (<10 cfu/g) in HPP
avocado puree and guacamole (689 MPa / 100,000 psi; 5
min; pH 4.3) for 30 days of storage at 5, 15 and 25 ºC (41,
59 and 77 ºF, respectively) (Palou et al., 2000).
Challenge tests
Besides controlling spoilage microorganisms, high pressure
processing has demonstrated high inactivation levels on
pathogenic bacteria. Guacamole was reported as a possible
vehicle in 136 outbreaks along United States from 1973 to
2008 (Kendall et al., 2013).
High hydrostatic pressure (600 MPa / 87,000 psi, 3 min)
inactivated efficiently L. monocytogenes (> 7 log cycles),
Salmonella spp. (> 5 log cycles) and E. coli (> 6 log cycles) in
avocado pulp and guacamole (Figure 2).
Figure 1. Total aerobic and lactic acid bacteria in HPP guacamole (600 MPa, 3
min) stored at 4 ºC (Jacobo-Velázquez and Hernández-Brenes, 2010).
Figure 2. HPP Inactivation of Listeria monocytogenes, Salmonella spp., and
Escherichia coli in guacamole (600 MPa, 3 min) (Hiperbaric, not published)

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Influence of processing parameters and product
characteristics
Microbial inactivation levels depend on the pressure and
holding time as well factors related to food product such as
water activity (aw) or pH, and environmental factors such
as temperature.
The lower water activity (aw) (or higher Brix degrees) the
lower effectiveness high pressure is (Goh et al., 2007);
therefore, the technology is very effective on guacamole
and avocado products, since aw is higher than 0.90.
The pH of a product is also a key factor to consider,
working in synergy with HPP: the lower pH a product has,
the more effective microbial inactivation by HPP is
reached.
Neetoo and Chen (2012) investigated the inactivation of
Salmonella spp. in guacamole. Peppers, an ingredient of
guacamole, were inoculated by five strains cocktail of this
pathogen. The storage time (0 or 24h, room temperature)
of the condiments prior to HPP (500 MPa / 72,520 psi, 2
min) as well as pH (4.3 - 5.3) and the type of acidulant
(vinegar and lemon juice) affected the inactivation levels of
Salmonella.
Total inactivation (i.e. no detection (<0.7 log cfu/g)) was
only reached at pH 4.3 (using vinegar or lemon juice), or
when vinegar was used as acidulant, and after 24h of
storage time at room temperature prior to HPP (Figure 3).
HPP does not inactivate bacterial spores (but mold spores
are inactivated). Regarding HACCP, HPP cannot be used to
control Clostridium botulinum or any other bacterial spore.
Products with a pH higher than 4.6 are at risk for spore
germination and therefore they must be kept refrigerated
for the entire life of the product. We recommend acidifying
HPP products below pH 4.6 whenever possible to prevent
spore germination.
SENSORIAL QUALITY AND ENZYMATIC
INACTIVATION
HPP is a suitable technology for controlling microbial
spoilage during long periods; however, when food products
are microbiologically stable, the end of their shelf life is
generally limited by changes in their sensory and
nutritional properties, which are mainly caused by
deteriorative biochemical/enzymatic reactions.
Figure 3. Survival of Salmonella spp.in different acidulated HPP guacamole
and different storage times (0h and 24h) prior pressurization (500 MPa, 2
min).*ND: not detectable. (Neetoo and Chen, 2012).

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These reactions are catalyzed by endogenous enzymes
such as polyphenol oxidase (PPO) and lipoxygenase (LOX).
These enzymes affect directly the components responsible
of color, flavor and texture of avocado products. When
pulp is removed from the intact fruit for processing, the
tissues undergo partial disruption of cellular organelles
releasing enzymes and their substrates causing
deterioration reactions.
Depending on type of enzyme, physicochemical
characteristics of the food product (pH, aw) and the
conditions of HPP process (pressure and holding time),
enzymes could be partly or moderately inactivated.
Storage under refrigeration (4 ºC / 39.2 ºF) minimizes
detrimental quality modifications induced by residual
activities (Jacobo-Velázquez and Hernández-Brenes, 2011).
Color
Enzymatic browning is the main change in appearance of
avocado products and it is catalyzed by PPO. Enzymatic
browning also depends on presence of O2 in contact of
food product. Vacuum and modified atmosphere packaging
allow controlling O2, slowing down browning. Both
packaging technologies can be used jointly to high pressure
technology.
HPP at 600 MPa (87,000 psi) for 3 min reduced
immediately around 50% PPO activity on avocado puree
(pH 6.35) (Jacobo-Velázquez and Hernández-Brenes, 2010).
Refrigeration is essential for controlling PPO activity in
guacamole and avocado products before and after HPP
processing in order to slow down the enzymatic reaction.
Although PPO activity reached a maximum peak at day 10,
the rest of storage time, residual PPO activity remained at
initial values, around 50% (Jacobo-Velázquez and
Hernández-Brenes, 2010; Figure 4).
Despite of PPO is partly deactivate, several studies have
shown that pressurized avocado paste does not present
significant color modification compared to freshly prepared
puree by using instrumental color measurements and
trained sensory panelists (López-Malo, et al., 1999; Palou
et al., 2000; Jacobo-Velázquez and Hernández-Brenes,
2010).
Factors affecting PPO inactivation
PPO inactivation does not depend only on HPP conditions,
but pH of the product plays a relevant role on enzyme
stability. López-Malo, et al. (1999) studied the effect of pH
on inactivation of PPO in avocado puree. At low pH values
and higher pressures for long holding time, the efficacy of
Figure 4. Residual enzymatic activity of PPO and LOX on pressurized (600 MPa,
3 min) avocado puree at refrigerated storage (Jacobo-Velázquez and
Hernández-Brenes, 2010).

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HPP on inactivating PPO is higher; residual activity of PPO
was below 30% (Figure 5).
Storage temperature plays a relevant role on maintaining
initial product color, decreasing rate of enzymatic
browning and other degradation reactions over natural
pigments. According to Palou et al. (2000), only HPP
guacamole stored at 5 ºC (41 ºF) retained green hues for
more than 10 days; higher storage temperatures (15 and
25 ºC / 59 and 77 ºF) led to changes in color.
Flavor
Activities of endogenous enzymes catalyze biochemical
reactions which lead to undesirable sensory quality traits.
Besides catalyzing product browning, PPO activity has also
associated with the formation of undesirable flavors
(Weemaes et al,. 1998). HPP is able to reduce partly PPO
activity, slowing down enzymatic browning and the
collateral undesirable effects, as it has been previously
shown.
In addition to PPO, LOX enzyme is contained in the pulp of
avocado. This enzyme degrades lipids and related
compounds such as carotenoids, generating low molecular
substances responsible of off-flavors and rancid volatile
compounds (Jacobo-Velázquez and Hernández-Brenes,
2010, 2011).
LOX is less stable to pressure than PPO. At commercial HPP
conditions (600 MPa / 87,000 psi, 3 min), residual LOX
activity in avocado puree (pH 6.35) is reduced to 45%
immediately after HPP compared to the control (Jacobo-
Velázquez and Hernández-Brenes, 2010; Jacobo-Velázquez
et al. 2012).
In HPP avocado puree, residual LOX activity during
refrigeration storage (4 ºC, 39.2 ºF) shows a similar
behavior than PPO (Figure 4). Residual activity reaches a
maximum peak at day 10, after that, LOX activity decreases
below 60% at the end of storage at day 45 (Jacobo-
Velázquez and Hernández-Brenes, 2010). Residual LOX
activity did not affect consumers’ acceptability of HPP
guacamole and rancid flavor was not developed (Jacobo-
Velázquez and Hernández-Brenes, 2011).
Overall sensory evaluation
Sensory evaluation of HPP guacamole using trained
panelists showed no significant differences in the different
sensory attributes compared to freshly-made products. The
results from Palou et al. (2000) showed HPP technology
preserve sensory quality of avocado products, a quite
sensitive food product.
Figure 5. Residual PPO activity in HPP avocado puree at different pressure
levels (for 20 min) and different puree pH values (López-Malo, et al., 1999).

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Enzymatic browning is a shelf-life limiting factor of
guacamole. Despite of HPP technology can not completely
deactivate PPO enzyme. Refrigeration and packaging were
a significant factor when a trained panel did not consider
the changes in color of HPP guacamole as a critical
descriptor (Jacobo-Velázquez and Hernández-Brenes,
2011).
NUTRIENTS RETENTION
It is well known that HPP technology has a minimal effect
on low molecular-weight compounds after pressurization,
contributing the conservation of sensitive molecules such
as vitamins, pigments, antioxidants and other substances
with relevant biological activity, sensitive to thermal
treatments (Jacobo-Velázquez and Hernández-Brenes,
2012).
Vegetables are one of the important sources of nutrients
and bioactive molecules in human diet. Lutein, vitamins C
and E, and persenone A and B are biological active
substances present in avocado pulp, protecting cells and
tissues from free radicals, inflammation and carcinogenesis
processes (Kim et al., 2000).
Other antioxidants and bioactive compounds present in
avocados are phenolics, monounsaturated fatty acids,
lycopene and other carotenoids such as xanthins. The
presence of these compounds is desired from the creation
of guacamole until before its consumption.
Carotenoids
These pigments are relevant in human physiology since
they have antioxidant and vitamin A activity. The
concentration of total carotenoids measured in HPP (600
MPa / 87,000 psi, 3 min) avocado puree was 2.3 times
higher than in unprocessed puree (Jacobo-Velázquez et al.,
2012). High pressure processing increased the measurable
content of the most of carotenoids in avocado puree
immediately after HPP compared to fresh avocado puree
(Relative concentration C/C0 = 1) (Figure 6)
Carotenoids are bound to proteins in the plant cell
membrane surface. Since high pressure induces changes in
protein structure, the complex protein-carotenoids may be
disassembled, facilitating the extraction of these
compounds from food matrices (Sánchez-Moreno et al.,
2003). Researchers stated the differences for individual
carotenoid can be attributed to the different degree of
association carotenoid-binding protein.
Figure 6. Relative concentration of total and individual carotenoids in HPP
(600 MPa, 3 min) avocado puree (where fresh-prepared avocado puree
composition has a relative concentration of 1.0 –red line-) at day 0 and day 40
at refrigerated storage (Jacobo-Velázquez et al., 2012)

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Increase in the extractability of carotenoids in HPP
guacamole during refrigerated storage may lead to a
positive impact on the bioavailability of these compounds;
although it has not been demonstrated from a
pharmacokinetic standpoint.
Although during refrigerated storage (day 40), the total
content of carotenoids decreased, it was still higher than
the content in unprocessed puree (Figure 6). Decrease in
carotenoids content during storage may be related to
degradation by residual LOX (Jacobo-Velázquez et al.,
2012).
Antioxidant activity
Additionally to determine how processing technologies
modify the concentration of the antioxidant species in food
products, it is also important to measure the effects of
processing on the overall antioxidant activity of the foods,
since it is related to the biological beneficial effects such as
preventing oxidative damage to important biomolecules
such as DNA, proteins and lipids.
HPP immediately decreases about 25% of the total
antioxidant activity (determined by ORAC method) in
avocado paste (Jacobo-Velázquez et al., 2012). However,
from day 5, avocado paste recovered its initial antioxidant
activity (Control values), reaching a maximum peak at day
10. After 15 days at refrigerated storage (4 ºC, 39.2 ºC),
antioxidant activity went back to control values (Figure 7).
According to Jacobo-Velázquez et al. (2012), the dynamics
of antioxidant activity during refrigerated storage is a
complex process: initial reduction due to enzymatic
degradation of vitamin C and oxidation of polyphenols by
residual PPO activity. After initial decrease, antioxidant
activity would increase since some antioxidants such as
carotenoids and phenolic compounds start to release from
tissues and cells affected by pressurization, reaching a
maximum value at day 15 (Figure 7).
Stabilization of total antioxidant activity might be due to a
balance between the releasing of compounds from the
interior of plant cells and degradation of antioxidants
caused by residual PPO and LOX enzymes.
Figure 7. Total antioxidant activity (µmol Trolox eq/ g dry weight) throughout
refrigerated storage of HPP (blue bars) avocado paste (600 MPa, 3 min)
compared to control (white bar) (Jacobo-Velázquez et al., 2012).

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CONCLUSIONS
Guacamole and avocado products have successfully been
processed by high hydrostatic pressure around the globe
since 1996. HPP technology increases the shelf-life of the
avocado products, maintaining, and in some cases
enhancing, their nutritional quality, while the products
keep the freshness and sensory attributes of home-made
product, avoiding the use of additives.
High Pressure Processing technology started its industrial
evolution hand in hand with the avocado processing
industry. You can see how avocado processing industry is
applying HPP technology on its products:
http://www.hiperbaric.com/en/avocado-products
You can get more information about Hiperbaric and high
pressure processing of foods on the next links:
http://www.hiperbaric.com
http://blog.hiperbaric.com/en/
You can also contact us via e-mail. We are pleased to
answer your questions.
info@hiperbaric.com

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